Pharmaceutical composition

ABSTRACT

A pharmaceutical composition comprising a tricyclic compound (I) or its pharmaceutically acceptable salt, an oil substance, a surfactant, a hydrophilic substance, water, and optionally a pH control agent, with enhanced stability, absorbability and/or a low irritation potential, is provided.

TECHNICAL FIELD

This invention relates to a pharmaceutical composition comprising atricyclic compound of general formula (I) or a pharmaceuticallyacceptable salt thereof which features stability, excellentabsorbability, and/or a reduced irritation potential. Thispharmaceutical composition is of value for the treatment or preventionof inflammatory or hyperproliferative skin diseases or cutaneousmanifestations of immunologically-mediated diseases.

BACKGROUND ART

The tricyclic compound (I) and its pharmaceutically acceptable salt foruse in accordance with this invention is known to have excellentimmunosuppressive activity, antimicrobial activity and otherpharmacological activities and, as such, be of value for the treatmentor prevention of rejection reactions by transplantation of organs ortissues, graft-vs.-host diseases, autoimmune diseases, and infectiousdiseases [Japanese Kokai Tokkyo Koho S61-1481B1, EP-A-0323042, etc.].

Particularly, those species of tricyclic compound (I) which aredesignated as FR900506 (=FK506 Substance), FR900520, FR900523, andFR900525 are products produced by microorganisms of the genusStreptomyces, such as Streptomyces tsukubaensis No. 9993 [deposited withNational Institute of Bioscience and Human Technology Agency ofIndustrial Science and Technology (formerly Fermentation ResearchInstitute Agency of Industrial Science and Technology ), at 1-3, Higashi1-chome, Tsukuba-shi, Ibaraki, Japan, date of deposit Oct. 5, 1984,accession number FERM BP-927] or Streptomyces hygroscopicus subsp.vakushimaensis No. 7238 [deposited with National Institute offBioscience and Human Technology Agency of Industrial Science andTechnology (formerly Fermentation Research Institute Agency ofIndustrial Science and Technology ), at 1-3, Higashi 1-chome,Tsukuba-shi, Ibaraki, Japan, date of deposit Jan. 12, 1985, accessionnumber FERM BP-928]. The FK506 Substance of the following chemicalformula, in particular, is a representative compound.

Chemical name:17-allyl-1,14-dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]octacos-18-ene-2,3,10, 16-tetraone

It has been demonstrated that the above-mentioned FK506 Substance hasquite excellent immunosuppressive activity and is useful for thetreatment or prevention of rejection by organ transplantation and thetreatment or prevention of diseases in the field of ophthalmology.

Japanese Kokai Tokkyo Koho H1-157913 discloses that a solution of FK506Substance in ethanol is effective in the suppression of inflammation andthat FK506 Substance can be formulated into a lotion, gel, or cream.However no specific dosage forms of the kinds are described.

Japanese Kokai Tokkyo Koho H5-17481 discloses an ointment comprising atricyclic compound (I) or its pharmaceutically acceptable salt, at leasta sufficient amount of a dissolution/absorption promoter to dissolve thesame, and an ointment base.

W094/28894 discloses a lotion comprising a tricyclic compound (I) or itspharmaceutically acceptable salt, a dissolution/absorption promoter, aliquid medium, and optionally an emulsifier and/or a thickening(rheology modifier).

Heretofore, ointments have mainly been used in the treatment of skindiseases. However, different dosage forms suited to different clinicalmanifestations and different application sites are being demanded.

The inventors of this invention have studied possible pharmaceuticalcompositions for compounds of general formula (I) inclusive of FK506Substance and found a dosage form having many desirable characteristicssuch as high stability, high transdermal absorbability, and/or reduceddermal irritancy. Thus, specifically this invention is directed to anhydrophilic semi-solid composition for external use containing saidtricyclic compound.

DISCLOSURE OF INVENTION

In accordance with this invention there is provided a pharmaceuticalcomposition comprising a tricyclic compound (I) or its pharmaceuticallyacceptable salt, an oil substance, a surfactant, a hydrophilicsubstance, and water, and further optionally a pH control agent.

The tricyclic compound for use in this invention can be expressed by thefollowing general formula (I).

(wherein each of adjacent pairs of R¹ and R², R³ and R⁴ or R⁵ and R⁶independently

(a) is two adjacent hydrogen atoms, or

(b) may form another bond formed between the carbon atoms to which theyare attached, and further, R² may be an alkyl group;

R⁷ is a hydrogen atom, a hydroxy group, a protected hydroxy group or analkoxy group, or an oxo group together with R¹;

each of R⁸ and R⁹ is independently a hydrogen atom or a hydroxy group;

R¹⁰ is a hydrogen atom, an alkyl group, an alkyl group substituted byone or more hydroxy groups, an alkenyl group, an alkenyl groupsubstituted by one or more hydroxy groups, or an alkyl group substitutedby an oxo group;

X is an oxo group, (a hydrogen atom and a hydroxy group) (a hydrogenatom and a hydrogen atom), or a group represented by the formula —CH₂O—;

Y is an oxo group, (a hydrogen atom and a hydroxy group) (a hydrogenatom and a hydrogen atom), or a group represented by the formulaN—NR¹¹R¹² or N—OR^(13;)

each of R¹¹ and R¹² is independently a hydrogen atom, an alkyl group, anaryl group or a tosyl group;

each of R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²² and R²³ is independentlya hydrogen atom or an alkyl group;

each of R²⁰ and R²¹ is independently an oxo group or (R²⁰a and ahydrogen atom) or (R²¹a and a hydrogen atom) in which each of R²⁰a andR²¹a is independently a hydroxy group, an alkoxy group or a grouprepresented by the formula —OCH₂OCH₂CH₂OCH₃, or R²¹a is a protectedhydroxy group, or R²⁰a and R²¹a may together represent an oxygen atom inan epoxide ring;

n is an integer of 1 or 2; and

in addition to the above definitions, Y, R¹⁰ and R²³, together with thecarbon atoms to which they are attached, may represent a saturated orunsaturated 5- or 6-membered nitrogen, sulfur and/or oxygen containingheterocyclic ring optionally substituted by one or more groups selectedfrom the group consisting of an alkyl, a hydroxy, an alkyl substitutedby one or more hydroxy groups, an alkoxy, a benzyl and a group of theformula —CH₂Se (C₆H₅)).

The above compound (I) or its pharmaceutically acceptable salt can beprovided by the same technology as that described in the two patentgazettes referred to above. Particularly, the tricyclic compoundsproduced by fermenting Streptomyces tsukubaensis No. 9993 (FERM BP-927)or Streptomyces hygroscopicus subsp. yakushimaensis No. 7238 (FERMBP-928) are known by the identification nos. of FR-900506, FR-900520,FR-900523, and FR-900525 (Japanese Kokai Tokkyo Koho S61-148181).

The various definitions given in the above general formula (I), genericand subgeneric examples thereof, and preferred species are now explainedand shown in detail.

The term “lower” means, unless otherwise indicated, a group having 1 to6 carbon atoms.

Preferable examples of the “alkyl groups” include a straight or branchedchain aliphatic hydrocarbon residue for example, a lower alkyl groupsuch as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl,neopentyl and hexyl.

Preferable examples of the “alkenyl groups” include a straight orbranched chain aliphatic hydrocarbon residue having one double-bond, forexample, a lower alkenyl group such as vinyl, propenyl (e.g., allylgroup), butenyl, methylpropenyl, pentenyl and hexenyl.

Preferable examples of the “aryl groups” include phenyl, tolyl, xylyl,cumenyl, mesityl and naphthyl.

Preferable protective groups in the “protected hydroxy groups” are1-(lower alkylthio) (lower) alkyl group such as a lower alkylthiomethylgroup (e.g., methylthiomethyl, ethylthiomethyl, propylthiomethyl,isopropylthiomethyl, butylthiomethyl, isobutylthiomethyl,hexylthiomethyl, etc.), more preferably C₁-C₄ alkylthiomethyl group,most preferably methylthiomethyl group;

trisubstituted silyl group such as a tri(lower)alkylsilyl (e.g.,trimethylsilyl, triethylsilyl, tributylsilyl, tert-butyldimethylsilyl,tri-tert-butylsilyl, etc.) or lower alkyl-diarylsilyl (e.g.,methyldiphenylsilyl, ethyldimethylsilyl, propyldiphenylsilyl,tert-butyldiphenyl-silyl, etc.), more preferably tri(C₁—C₄)alkylsilylgroup and C₁-C₄ alkyldiphenylsilyl group, most preferablytert-butyldimethylsilyl group and tert-butyldiphenylsilyl group; and anacyl group such as an aliphatic, aromatic acyl group or an aliphaticacyl group substituted by an aromatic group, which are derived from acarboxylic acid, sulfonic acid or carbamic acid.

Examples of the aliphatic acyl groups include a lower alkanoyl groupoptionally having one or more suitable substituents such as carboxy,e.g., formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl,isovaleryl, pivaloyl, hexanoyl, carboxyacetyl, carboxypropionyl,carboxybutyryl, arboxyhexanoyl, etc.; acyclo(lower)alkoxy(lower)alkanoyl group optionally having one or moresuitable substituents such as lower alkyl, e.g., cyclopropyloxyacetyl,cyclobutyloxypropionyl, cycloheptyloxybutyryl, menthyloxyacetyl,menthyloxypropionyl, menthyloxybutyryl, menthyloxypentanoyl,menthyloxyhexanoyl, etc.; a camphorsulfonyl group; or a loweralkylcarbamoyl group having one or more suitable substituents such ascarboxy or protected carboxy, for example, carboxy(lower)alkylcarbamoylgroup (e.g., carboxymethylcarbamoyl, carboxyethylcarbamoyl,carboxypropylcarbamoyl, carboxybutylcarbamoyl, carboxypentylcarbamoyl,carboxyhexylcarbamoyl, etc.),tri-lower)alkylsilyl(lower)alkoxycarbonyl(lower)-alkylcarbamoyl group(e.g., trimethylsilylmethoxy-carbonylethylcarbamoyl,trimethylsilylethoxycarbonylpropylcarbamoyl,triethylsilylethoxycarbonylpropylcarbamoyl,tert-butyldimethylsilylethoxycarbonylpropylcarbamoyl,tri-methylsilylpropoxycarbonylbutylcarbamoyl, etc.) and so on.

Examples of the aromatic acyl groups include an aroyl group optionallyhaving one or more suitable substituents such as nitro, e.g., benzoyl,toluoyl, xyloyl, naphthoyl, nitrobenzoyl, dinitrobenzoyl,nitronaphthoyl, etc.; and an arenesulfonyl group optionally having oneor more suitable substituents such as halogen, e.g., benzenesulfonyl,toluenesulfonyl, xylenesulfonyl, naphthalenesulfonyl,fluorobenzenesulfonyl, chlorobenzenesulfonyl, bromobenzenesulfonyl,iodobenzenesulfonyl, etc.

Examples of the aliphatic acyl groups substituted by an aromatic groupinclude ar(lower)alkanoyl group optionally having one or more suitablesubstituents such as lower alkoxy or trihalo(lower)alkyl, e.g.,phenylacetyl, phenylpropionyl, phenylbutyryl,2-trifluoromethyl-2-methoxy-2-phenylacetyl,2-ethyl-2-trifluoromethyl-2-phenylacetyl,2-trifluoromethyl-2-propoxy-2-phenylacetyl, etc.

More preferable acyl groups among the aforesaid acyl groups are C₁-C₄alkanoyl group optionally having carboxy,cyclo(C₅-C₆)alkoxy(C₁-C₄)alkanoyl group having two (C₁-C₄) alkyls at thecycloalkyl moiety, camphorsulfonyl group, carboxy(C₁-C₄)alkylcarbamoylgroup, tri(C₁-C₄)alkylsilyl(C₁-C₄)alkoxycarbonyl(C₁-C₄)alkylcarbamoylgroup, benzoyl group optionally having one or two nitro groups,benzenesulfonyl group having halogen, orphenyl(C₁-C₄)alkanoyl grouphaving C₁-C₄ alkoxy and trihalo(C₁-C₄)alkyl group. Among these, the mostpreferable ones are acetyl, carboxypropionyl, menthyloxyacetyl,camphorsulfonyl, benzoyl, nitrobenzoyl, dinitrobenzoyl,iodobenzenesulfonyl and 2-trifluoromethyl-2-methoxy-2-phenylacetyl.

Preferable examples of the “5- or 6-membered nitrogen, sulfur and/oroxygen containing heterocyclic ring” include a pyrrolyl group and atetrahydrofuryl group.

The pharmaceutically acceptable salt of the compound (I) includesconventional non-toxic and pharmaceutically acceptable salt such as thesalt with inorganic or organic bases, specifically, an alkali metal saltsuch as sodium salt and potassium salt, an alkali earth metal salt suchas calcium salt and magnesium salt, an ammonium salt and an amine saltsuch as triethylamine salt and N-benzyl-N-methylamine salt.

With respect to the compound (I), it is to be understood that there maybe conformers and one or more stereoisomers such as optical andgeometrical isomers due to asymmetric carbon atom(s) or double bond(s),and such conformers and isomers are also included within the scope ofthe present invention.

The compound of the formula (I) or its pharmaceutically acceptable saltcan be in the form of a solvate, which is included within the scope ofthe present invention. The solvate preferably include a hydrate and anethanolate.

The preferred examples of the tricyclic compound (I) is the one, whereineach of adjacent pairs of R³ and R⁴ or R⁵ and R⁶ independently formanother bond formed between the carbon atoms to which they are attached;

each of R⁸ and R²³ is independently a hydrogen atom;

R⁹ is a hydroxy group;

R¹⁰ is a methyl group, an ethyl group, a propyl group or an allyl group;

X is (a hydrogen atom and a hydrogen atom) or an oxo group;

Y is an oxo group;

each of R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, and R²² is a methyl group;

each of R²⁰ and R²¹ is independently (R²⁰a and a hydrogen atom) or (R²¹aand a hydrogen atom) in which each of R²⁰a and R²¹a is a hydroxy groupor an alkoxy group, or R²¹a is a protected hydroxy group; and

n is an integer of 1 or 2.

FK506 Substance is the most preferable compound belonging to thetricyclic compound (I). Other preferable compounds are listedhereinbelow.

1,14-Dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy-13,17,19,21,27-pentamethyl-11,28-dioxa-4-azatricyclo[(22.3.1.0^(4,9)]octacos-18-ene-2,3,10,16-tetraone,

12-[2-(4-acetoxy-3-methoxycyclohexyl)-1-methylvinyl]-17-allyl-1,14-dihydroxy-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0_(4,9)]octacos-18-ene-2,3,10,16-tetraone,

17-allyl-1,14-dihydroxy-23,25-dimethoxy-13,19,21,27-tetramethyl-12-[2-[4-(3,5-dinitrobenzoyloxy)-3-methoxycyclo-hexyl]-1-methylvinyl]-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]-octacos-18-ene-2,3,10,16-tetraone,

17-allyl-12-[2-[4-[(−)-2-trifluoromethyl-2-methoxy-2-phenylacetoxy]-3-methoxycyclohexyl]-1-methylvinyl]-1,14-dihydroxy-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0_(4,9)]octacos-18-ene-2,3,10,16-tetraone.

17-ethyl-1,14-dihydroxy-12-[2-(4-hydroxy-3-methoxycyclo-hexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]octacos-18-ene-2,3,10,16-tetraone(FR900520), and

17-ethyl-1,14,20-trihydroxy-12-[2-(3,4-dihydroxycyclo-hexyl)-1-methylvinyl-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo(22.3.1.0_(4,9)]octacos-18-ene-2,3,10,16-etraone.

The oil substance for use in this invention need only to be a substancecapable of dissolving tricyclic compound (I) or its pharmaceuticallyacceptable salt. Preferable ones are, for example, fatty acid esters andalcohols.

Monohydric alcohol fatty acid esters (isopropyl myristate, ethylmyristate, butyl myristate, isocetyl myristate, octyldodecyl myristate,isopropyl palmitate, isostearyl palmitate, isopropyl isostearate,isocetyl isostearate, butyl stearate, isocetyl stearate, cetylisooctanoate, ethyl linoleate, isopropyl linoleate, hexyl laurate, ethyloleate, decyl oleate, oleyl oleate, octyl dodecyl myristate, hexyl decyldimethyloctanoate, octyl dodecyl neodecanoate, etc.)

Dibasic acid diesters (diisopropyl adipate, dimethyl adipate, diethyladipate, diisobutyl adipate, diethyl sebacate, diisopropyl sebacate,dipropyl sebacate, diethyl phthalate, diethyl pimelate, etc.)

Alcohols (oleyl alcohol, cetanol, stearyl alcohol, 2-octyldodecanol,etc.)

In this invention the above-mentioned oil substances can be usedindependently or in a combination of two or more species.

Particularly from the standpoint of stability of the dosage form,absorbability and/or irritation potential, it is preferable, in manyinstances, to use a plurality of such oil substances in combination. Thepreferred combination may for example be a mixture of a monohydricalcohol fatty acid ester (e.g. isopropyl myristate) and a dibasic aciddiester (e.g. diethyl sebacate) in a suitable weight ratio (e.g.0.1˜10:1 (w/w), preferably 0.5˜2:1 (w/w)) . The most preferable weightratio thereof is 1:1 (w/w).

The proportion of the oil substance in the pharmaceutical composition ispreferably 2˜50% (w/w), more preferably 10˜40%, and most preferably20˜30%.

The surfactant for use in this invention is now described.

The surfactant that can be used includes pharmaceutically acceptableionic or nonionic surfactants but is preferably a nonionic surfactantwith an HLB number of not less than 10. More preferably, the followingether series and ester series surfactants can be mentioned.

Ethers

Polyoxyethylene alkyl ethers (polyoxyethylene oleyl ether,polyoxyethylene stearyl ether, polyoxyethylene cetyl ether,polyoxyethylene lauryl ether (Lauromacrogol J.P.), polyoxyethylenebehenyl ether, etc.)

Esters

Polyoxyethylene sorbitan fatty acid esters Polyoxyethylene sorbitanmonostearate, polyoxyethylene sorbitan monooleate, etc. (e.g. Tween 20,Tween 40, Tween 60, Tween 65, Tween 80, etc., all trademarks)

Polyethylene glycol fatty acid esters (polyethylene glycol monooleate,polyethylene glycol monostearate (e.g. polyoxyl stearate 40 J.P.),polyethylene glycol monolaurate, etc.)

Pentaglycerol fatty acid esters Pentaglycerol monolaurate, pentaglycerolmonomyristate, pentaglycerol monooleate, pentaglycerol monostearate,etc.

Glycerol fatty acid esters Glyceryl monostearate etc.

In working this invention, the above-mentioned surfactants can be usedindependently or in a combination of two or more species. The proportionof the surfactant in the pharmaceutical composition is preferably0.1˜15% (w/w) and more preferably 0.5˜5%(w/w).

The hydrophilic substance for use in this invention may be any substancethat is pharmaceutically acceptable and capable of imparting viscosityto liquids, thus including the following organic or inorganichydrophilic substances.

(1) Organic Substances

{circle around (1)} Natural polymers . . . gum arabic, guar gum,carrageenin, gum tragacanth, pectin, starch, gum xanthan, gelatin,casein, dextrin, cellulose

{circle around (2)} Semi-synthetic polymers . . . methylcellulose,ethylcellulose, hydroxyethylcellulose, hydroxy-propylcellulose,carboxymethylcellulose sodium, carboxymethylcellulose calcium,carboxymethylstarch, sodium alginate, propylene glycol alginate,

{circle around (3)} Synthetic polymers . . . carboxyvinyl polymer(Carbopol) polyvinyl alcohol, polyvinylpyrrolidone, polyethylene glycol,polyvinyl methyl ether, sodium polyacrylate

(2) Inorganic substances Bentonite, synthetic magnesium silicate,magnesium aluminosilicate, silicon oxide, etc.

The proportion of the hydrophilic substance in the pharmaceuticalcomposition of this invention is selected according to the desiredviscosity of the pharmaceutical composition and is preferably 0.1˜10%(wlw) and more-preferably 0.5˜2%.

Furthermore, in the instant invention, the pharmaceutical composition ispreferably maintained at a constant pH from safety points of view.Therefore, a pH control agent such as a buffer, an aqueous solution ofsodium hydroxide, or the like can be added in a suitable amount. The pHrange is preferably 3.5˜6 and more preferably 4˜5.

Where necessary, in addition to the above ingredients, the conventionalexcipient (e.g. petrolatum, propylene glycol, etc.), stabilizer(antioxidant etc.), coloring agent, sweetener, flavor, diluent,antiseptic (e.g. parahydroxybenzoates, benzalkonium chloride, sorbicacid, etc.), and other drug substances effective against diseases of theskin can be added.

Meanwhile, the pharmaceutical composition of this invention can beproduced by a process comprising the following steps.

(1) A step which comprises preparing a solution composed of saidtricyclic compound (I) or pharmaceutically acceptable salt thereof, oilsubstance, and surfactant;

(2) a step which comprises mixing the solution with water to give anemulsion; and

(3) a step which comprises mixing the emulsion with said hydrophilicsubstance, and further optionally pH control agent, with stirring. As analternative, this step may comprise mixing said hydrophilic substance,and further optionally pH control agent, with water and, then, mixingthe premix with the emulsion prepared as above with stirring.

The step (1) is preferably carried out at elevated temperature, forexample at 50˜90° C., preferably 60˜80 ° C.

In step (2), the water for use is also preferably heated ahead of timeto a temperature close to that of the solution prepared in step (1), andthe resulting emulsion is preferably cooled to a suitable temperature,for example 30° C., before step (3) is carried out.

The dosage of tricyclic compound (I) or its pharmaceutically acceptablesalt depends on the individual patient's age and the type and severityof disease but the usual daily therapeutic dose is about 0.001˜1000 mg,preferably about 0.005˜500 mg, and more preferably about 0.01˜100 mg, asthe active ingredient. Generally, an average of about 0.01 mg, 0.05 mg,0.1 mg, 0.5 mg, 1 mg, 5 mg, 10 mg, 25 mg, or 50 mg per dose isadministered.

The recommended proportion of tricyclic compound (I) in thepharmaceutical composition is 0.001˜20% (w/w) preferably 0.01˜10% (w/w),of the total composition.

EXAMPLES

The following examples illustrate the present invention in furtherdetail, it being to be understood that those examples are not intendedto limit the scope of the invention.

Example 1

The following ingredients (a), (b), and (c) were first admixed at 60˜80°C. to prepare a solution, to which (d) previously warmed to 60˜80° C.was added. The mixture was evenly emulsified with a homomixer and theresulting emulsion was cooled to 30° C. Then, (e) was added to theemulsion and the whole mixture was stirred to provide a pharmaceuticalcomposition in the form of an easily spreadable cream [formulation (1)].

Formulation (1) (% w/w) (a) FK506 Substance 0.1 (b) Isopropyl myristate25.0  (c) Polyoxyethylene [5.5] cetyl ether 5.0 (d) Purified water 68.9 (e) Carbopol 940 1.0

Example 2

A pharmaceutical composition (formulation (2)) was prepared according toa similar manner to that of Example 1, except that, after addition of(e), the mixture was adjusted to pH about 4.0 with a suitable amount of1N-aqueous solution of NaOH (f).

Example 3

Pharmaceutical compositions (formulations (3) and (4)) were preparedaccording to a similar manner to that of Examples 1 and 2.

Formulation (3) Formulation (4) (% w/w) (% w/w) (a) FK506 Substance 0.10.1 (b) Isopropyl myristate 25.0  25.0  (c) Polyoxyethylene [7] cetylether 5.0 — (c) Polyoxyethylene [10] oleyl ether — 5.0 (d) Purifiedwater 68.9  68.9  (e) Carbopol 940 1.0 1.0 (f) 1N NaOH aq. sol. q.s.q.s.

Example 4

Pharmaceutical compositions (formulations (5)-(8)) were preparedaccording to a similar manner to that of Examples 1 and 2.

Formulation (5) (6) (7) (8) (% w/w) (% w/w) (% w/w) (% w/w) (a) FK506Substance 0.1 0.1 0.1 0.1 (b) Isopropyl 25.0  25.0  25.0  25.0 myristate (c) Pentaglycerol 5.0 — — — monolaurate (c) Pentaglycerol —5.0 — — monomyristate (c) Pentaglycerol — — 5.0 — monooleate (c)Pentaglycerol — — — 5.0 monostearate (d) Purified water 68.9  68.9 68.9  68.9  (e) Carbopol 940 1.0 1.0 1.0 1.0 (f) 1N-NaOH aq. sol. q.s.q.s. q.s. q.s.

Example 5

Pharmaceutical compositions (formulations (9)-(12)) were preparedaccording to a similar manner to that of Examples 1 and 2.

Formulation (9) (10) (11) (12) (% w/w) (% w/w) (% w/w) (% w/w) (a) FK506Substance 0.1 0.1 0.1 0.1 (b) Isopropyl 25.0  25.0  25.0  25.0 myristate (c) Glycerin 5.0 — — — monostearate (c) Polyethylene — 5.0 — —glycol monostearate (c) Polyoxyethylene- — — 5.0 — [20] sorbitanmonostearate (c) Polyoxyethylene- — — — 5.0 [20] sorbitan monooleate (d)Purified water 68.9  68.9  68.9  68.9  (e) Carbopol 940 1.0 1.0 1.0 1.0(f) 1N-NaOH aq. sol. q.s. q.s. q.s. q.s.

Example 6

Pharmaceutical compositions (formulations (13)-16)) were preparedaccording to a similar manner to that of Examples 1 and 2.

Formulation (13) (14) (15) (16) (% w/w) (% w/w) (% w/w) (% w/w) (a)FK506 Substance 0.1 0.1 0.1 0.1 (b) Diethyl sebacate 25.0  25.0  — 25.0 (b) Oleyl alcohol — — 25.0  — (c) Polyoxyethylene- 5.0 — — — [10]behenyl ether (c) Polyoxyethylene- — 5.0 — — [5.5] cetyl ether (c)Polyoxyethylene- — — 5.0 — [21] lauryl ether (c) Polyoxyethylene — — —5.0 [20] sorbitan monooleate (d) Purified water 68.9  68.9  68.9  68.9 (e) Carbopol 940 1.0 1.0 1.0 1.0 (f) 1N-NaOH aq. sol. q.s. q.s. q.s.q.s.

Example 7

The following ingredients (a), (b), and (c) were admixed at 60˜80° C. toprepare a solution, to which (d₁) preheated to 60˜80° C. was added. Themixture was evenly homogenized with a homomixer and, then, cooled to 30° C.

To this emulsion was added a gel prepared from (e) (d₂), and (f) and themixture was stirred thoroughly to provide a pharmaceutical compositionin the form of a well-spreadable cream [formulation (17)].

Formulation (17) (% w/w) (a) FK506 Substance 0.1 (b) Isopropyl myristate25.0  (c) Polyoxyethylene [5.5] cetyl ether 5.0 (d₁) Purified water34.4  (e) Carbopol 940 1.0 (d₂) Purified water 34.5  (f) 1N-NaO aq. sol.q.s.

Example 8

A pharmaceutical composition (formulation (18)) was prepared accordingto a similar manner to that of Example 7.

Formulation (18) (% w/w) (a) FK506 Substance 0.1 (b) Diethyl sebacate25.0 (c) Polyoxyethylene [20] sorbitan monooleate  5.0 (d₁) Purifiedwater 34.4 (e) Carbopol 940  1.0 (d₂) Purified water 34.5 (f) 1N-NaOHaq. sol. q.s.

Example 9

Pharmaceutical compositions (formulations (19)-(21)) were preparedaccording to a similar manner to that of Example 7.

Formulation (19) (20) (21) (% w/w) (% w/w) (% w/w) (a) FK506 Substance0.03 0.1 0.3 (b₁) Isopropyl myristate 20 20 20 (b₂) Diethyl sebacate 2020 20 (c) polyoxyethylene [20] 2.5 2.5 2.5 sorbitan monooleate (d₁) +(d₂) Purified water 56.47 56.4 56.2 (e) Carbopol 940 1.0 1.0 1.0 (f)1N-NaOH aq. sol. q.s. q.s. q.s.

Example 10

The protocol and results of a transdermal absorbability experiment usingpharmaceutical compositions of the present invention are describedbelow.

Transdermal Absorbability Test

Using the formulations 2, 3, 7, 12, and 20 prepared in the foregoingexamples, an in vivo transdermal absorbability experiment was performed.

As experimental animals, 3 male 7-week-old SD rats were used. With therat in supine position on a stereotactic apparatus, the hair coat in theabdominal region was clipped off with an electric clipper and, then, adepilatory cream (Eva-Cream, manufactured by Tokyo TanabePharmaceutical) was applied. The skin area thus treated was washed withwater 10 minutes after application of the cream for depilation. The ratwas returned to its cage and allowed to rest for 24 hours. Thereafter,the rat was in supine position on the stereotactic apparatus, arectangular area measuring 2.5 cm×4 cm was marked off at four corners onthe depilated abdominal area of the rat and 50 mg of the test sample wasapplied over the square. At 1, 3, 5, 8, and 24 hours after thisapplication, 0.3 ml of blood was drawn from the subclavian vein with anEDTA-pretreated syringe. After thorough mixing with EDTA, the blood wasstored frozen until assayed.

Using the above blood samples, the whole blood FK506 Substanceconcentration was determined by enzyme-linked immunosorbent assay usingperoxidase as the enzyme (e.g. shown in Japanese Kokai Tokkyo KohoH1-92659).

The transdermal absorption parameter was calculated for each testsample. The results are shown in Table 1. In Table 1, AUC [0-24 hr]represents the area under the blood concentration-time curve over 0-24hours after application.

TABLE 1 Transdermal absorption parameter (n = 3, mean ± S.E.) AUC [0-24hr] Test sample (ng.hr/ml) Formulation 2 >30 Formulation 3 >30Formulation 7 >30 Formulation 12 >30 Formulation 20 >30

Effects of the Invention

In accordance with this invention there can be provided a pharmaceuticalcomposition of tricyclic compound (I), particularly a hydrophilicsemi-solid composition for external application, which is stable, easyto use, acceptable in feeling in use, and with a low irritationpotential and/or improved dermal penetration.

The pharmaceutical composition of the present invention is useful forthe treatment or prevention of Inflammatory or hyperproliferative skindiseases or cutaneous manifestations of immunologically-mediateddiseases (e.g. psoriasis, atopic dermatitis, contact dermatitis,eczematoid dermatitis, seborrheic dermatitis, lichen planus, pemphigus,bullous pemphigoid, epidermolysis bullosa, urticaria, angioedema,vasculitides, erythema, dermal eosinophilia, lupus erythematosus, acne,and alopecia areata) because of the pharmacologic activities possessedby the tricyclic compound (I)

Furthermore, the active ingredient, tricyclic compound (I), used in thepharmaceutical composition is useful for the therapy or prophylaxis ofthe following diseases.

Rejection reactions by transplantation of organs or tissues such as theheart, kidney, liver, bone marrow, skin, cornea, lung, pancreas, smallintestine, limb, muscle, nerve, intervertebral disc, trachea, myoblast,cartilage, etc.;

graft-versus-host reactions following bone marrow transplantation;

autoimmune diseases such as rheumatoid arthritis, systemic lupuserythematosus, Hashimoto's thyroiditis, multiple sclerosis, myastheniagravis, type I diabetes, etc.;

and infections caused by pathogenic microorganisms (e.g. Aspergillusfumigatus, Fusarium oxysporum, Trichophyton asteroides, etc.);

autoimmune diseases of the eye (e.g. keratoconjunctivitis, vernalconjunctivitis, uveitis associated with Behcet's disease, keratitis,herpetic keratitis, conical keratitis, corneal epithelial dystrophy,keratoleukoma, ocular premphigus, Mooren's ulcer, scleritis, Graves'ophthalmopathy, Vogt-Koyanagi-Harada syndrome, keratoconjunctivitissicca (dry eye), phlyctenule, iridocyclitis, sarcoidosis, endocrineophthalmopathy, etc.);

reversible obstructive airways diseases [asthma (e.g. bronchial asthma,allergic asthma, intrinsic asthma, extrinsic asthma, and dust asthma),particularly chronic or inveterate asthma (e.g. late asthma and airwayhyper-responsiveness) bronchitis, etc.];

mucosal or vascular inflammations (e.g. gastric ulcer, ischemic orthrombotic vascular injury, ischemic bowel diseases, enteritis,necrotizing enterocolitis, intestinal damages associated with thermalburns, leukotriene B4-mediated diseases);

intestinal inflammations/allergies (e.g. coeliac diseases, proctitis,eosinophilic gastroenteritis, mastocytosis, Crohn's disease andulcerative colitis); food-related allergic diseases with symptomaticmanifestation remote from the gastrointestinal tract (e.g. migrain,rhinitis and eczema);

renal diseases (e.g. intestitial nephritis, Goodpasture's syndrome,hemolytic uremic syndrome, and diabetic nephropathy); nervous diseases(e.g. multiple myositis, Guillain-Barre syndrome, Meniere's disease,multiple neuritis, solitary neuritis, cerebral infarction, Alzheimer'sdisease, Parkinson's disease,amyotrophiclateralsclerosis(ALS) andradiculopathy);

cerebral ischemic disease (e.g., head injury, hemorrhage in brain (e.g.,subarachnoid hemorrhage, intracerebral hemorrhage), cerebral thrombosis,cerebral embolism, cardiac arrest, stroke, transient ischemic attack(TIA), hypertensive encephalopathy)

endocrine diseases (e.g. hyperthyroidism, and Basedow's disease);

hematic diseases (e.g. pure red cell aplasia, aplastic anemia,hypoplastic anemia, idiopathic thrombocytopenic purpura, autoimmunehemolytic anemia, agranulocytosis, pernicious anemia, megaloblasticanemia, and anerythroplasia);

bone diseases (e.g. osteoporosis);

respiratory diseases (e.g. sarcoidosis, pulmonary fibrosis, andidiopathic interstitial pneumonia);

skin diseases (e.g. dermatomyositis, leukoderma vulgaris, ichthyosisvulgaris, photosensitivity, and cutaneous T-cell lymphoma);

circulatory diseases (e.g. arteriosclerosis, atherosclerosis, aortitissyndrome, polyarteritis nodosa, and myocardosis);

collagen diseases (e.g. scleroderma, Wegener's granuloma, and Sjogren'ssyndrome);

adiposis;

eosinophilic fasciitis;

periodontal diseases (e.g. damage to gingiva, periodontium, alveolarbone or substantia ossea dentis);

nephrotic syndrome (e.g. glomerulonephritis);

male pattern alopecia, alopecia senile;

muscular dystrophy;

pyoderma and Sezary syndrome;

Addison's disease;

chromosome abnormality-associated diseases (e.g. Down's syndrome);

active oxygen-mediated diseases [e.g. organ injury (e.g. ischemiccirculation disorders of organs (e.g. heart, liver, kidney, digestivetract, etc.) associated with preservation, transplantation, or ischemicdiseases (e.g. thrombosis, cardial infarction, etc.));

intestinal diseases (e.g. endotoxin shock, pseudomembranous colitis, anddrug- or radiation-induced colitis): renal diseases (e.g. ischemic acuterenal insufficiency, chronic renal failure);

pulmonary diseases (e.g toxicosis caused by pulmonary oxygen or drugs(e.g. paracort, bleomycin, etc.), lung cancer, and pulmonary emphysema);

ocular diseases (e.g. cataracta, iron-storage disease (siderosis bulbi),retinitis, pigmentosa, senile plaques, vitreous scarring, corneal alkaliburn);

dermatitis (e.g. erythema multiforme, linear immunoglobulin A bullousdermatitis, cement dermatitis);

and other diseases (e.g. gingivitis, periodontitis, sepsis,pancreatitis, and diseases caused by environmental pollution (e.g. airpollution),aging, carcinogen, metastasis of carcinoma, andhypobaropathy)];

diseases caused by histamine release or leukotriene C4 release;restenosis of coronary artery following angioplasty and prevention ofpostsurgical adhesions;

Autoimmune diseases and inflammatory conditions (e.g., primary mucosaledema, autoimmune atrophic gastritis, premature menopause, malesterility, juvenile diabetes mellitus, pemphigus vulgaris, pemphigoid,sympathetic ophthalmitis, lens-induced uveitis, idiopathic leukopenia,active chronic hepatitis, idiopathic cirrhosis, discoid lupuserythematosus, autoimmune orchitis, arthritis (e.g. arthritisdeformans), or polychondritis);

Human Immunodeficiency Virus (HIV) infections AIDS;

allergic conjunctivitis;

hypertrophic cicatrix or keloid due to trauma, burn, or surgery.

In addition, the tricyclic compound (I) has liver regenerating activityand/or activities of stimulating hypertrophy and hyperplasia ofhepatocytes.

Therefore, the pharmaceutical composition of the present invention isuseful for the therapy or prophylaxis of liver diseases [e.g.immunogenic diseases (e.g. chronic autoimmune liver diseases such asautoimmune hepatic diseases, primary biliary cirrhosis or sclerosingcholangitis), partial liver resection, acute liver necrosis (e.g.necrosis caused by toxins, viral hepatitis, shock, or anoxia), hepatitisB, non-A non-B hepatitis, hepatocirrhosis, and hepatic failure (e.g.fulminant hepatitis, late-onset hepatitis and “acute-on-chronic” liverfailure (acute liver failure on chronic liver diseases))].

And further, the present composition is useful for preventing ortreating various diseases because of its useful pharmacological activitysuch as augmenting activity of chemotherapeutic effect, activity ofcytomegalovirus infection, anti-inflammatory activity, inhibitingactivity against peptidyl-prolyl isomerase or rotamase, antimalarialactivity, antitumor activity, and so on.

Among various formulations for the pharmaceutical composition of thisinvention, those formulations of reduced dermal irritancy are of valuefor treating or preventing atopic and other diseases of the skin, whilethose formulations with high dermal absorbability are particularlyuseful for treating or preventing psoriasis and other diseases of theskin.

What is claimed is:
 1. A pharmaceutical composition, comprising: (A) atricyclic compound of Formula (I) or a pharmaceutically acceptable saltof (A):

wherein each of adjacent pairs of R¹ and R², R³ and R⁴ or R⁵ and R⁶independently (a) are two adjacent hydrogen atoms, or (b) may formanother bond between the carbon atoms to which they are attached; R² isan alkyl group; R⁷ is a hydrogen atom, a hydroxy group, a protectedhydroxy group or alkoxy group, or an oxo group together with R¹; each ofR⁸ and R⁹ is independently a hydrogen atom or a hydroxy group; R¹⁰ is ahydrogen atom, an alkyl group, an alkyl group substituted by one or morehydroxy groups, an alkenyl group, an alkenyl group substituted by one ormore hydroxy groups, or an alkyl group substituted by an oxo group; X isan oxo group, (a hydrogen atom and a hydroxy group), two hydrogen atoms,or a group of formula -CH₂O-; Y is an oxo group, (a hydrogen atom and ahydroxy group), two hydrogen atoms, or a group of formulas N—NR¹¹R¹² orN—OR¹³; each of R¹¹ and R¹² is independently a hydrogen atom, an alkylgroup, an aryl group or a tosyl group; each of R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷,R¹⁸, R¹⁹, R²² and R²³ is independently a hyd or an alkyl group; each ofR²⁰ and R²¹ is independently an oxo group, (R²⁰a and a hydrogen atom),or (R²¹a and a hydrogen atom); wherein each of R²⁰a and R²¹a isindependently a hydroxy group, an alkoxy group or a group of formula—OCH₂OCH₂CH₂OCH₃, or R²¹a is a protected hydroxy group, or R²⁰a and R²¹aboth represent an oxygen atom in an epoxide ring; n is an integer of 1or 2; and Y, R¹⁰ and R²³, together with the carbon atoms to which theyare attached, may represent a saturated or unsaturated 5- or 6-memberednitrogen, sulfur and/or oxygen containing heterocyclic ring optionallysubstituted by at least one group selected from the group consisting ofan alkyl, a hydroxy, an alkyl substituted by at least one hydroxy group,an alkoxy, a benzyl and a group of the formula —CH₂Se(C₆H₅); (B) an oilsubstance combination of isopropyl myristate and diethyl sebacate; (C) asurfactant which is selected from the group consisting of apolyoxyethylene alkyl ether, a polyoxyethylene sorbitan fatty acidester, a pentaglycerol fatty acid ester and a glycerol fatty acid ester;(D) a hydrophilic substance which is capable of imparting viscosity toliquids; (E) water; and (F) optionally a pH control agent.
 2. Thepharmaceutical composition according to claim 1, wherein an amount ofsaid surfactant, said oil substance and said hydrophilic substance is0.1˜15% by weight, 2˜50% by weight, and 0.1˜10% by weight, respectively,based on the total weight of said composition.
 3. The pharmaceuticalcomposition according to claim 1, wherein a pH of said pharmaceuticalcomposition is 3.5˜6.
 4. The pharmaceutical composition according toclaim 1, wherein said hydrophilic substance is a carboxyvinyl polymer.5. The pharmaceutical composition according to claim 1, wherein saidsurfactant is a polyoxyethylene sorbitan fatty acid ester.
 6. Thepharmaceutical composition according to claim 1, wherein in saidtricyclic compound of Formula (I) each of said adjacent pairs of R³ andR⁴ or R⁵ and R⁶ independently may form another bond formed between thecarbon atoms to which they are attached; each of R⁸ and R²³ isindependently a hydrogen atom; R⁹ is a hydroxy group; R¹⁰ is a methylgroup, an ethyl group, a propyl group or an allyl group; X is a hydrogenatom and a hydrogen atom or an oxo group; Y is an oxo group; each of R⁴,R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, and R²² is a methyl group; each of R²⁰ and R²¹is independently (R²⁰a and a hydrogen atom) or (R²¹a and a hydrogenatom); wherein each of R²⁰a and R²¹a is a hydroxy group or an alkoxygroup, or R²¹a is a protected hydroxy group; and n is an integer of 1 or2.
 7. The pharmaceutical composition according to claim 6, wherein saidtricyclic compound of Formula (I) is17-allyl-1,14-dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]octacos-18-ene-2,3,10,16-tetraone.8. The pharmaceutical composition according to claim 7, wherein a pH ofsaid pharmaceutical composition is about 4-5.
 9. A process for producingthe pharmaceutical composition claimed in claim 1, comprising: mixing asolution comprising said tricyclic compound of Formula (I) or saidpharmaceutically acceptable salt of (A), said oil substance and saidsurfactant with water to prepare an emulsion; and mixing said emulsionwith said hydrophilic substance and optionally said pH control agent.10. The process according to claim 9, comprising: separately mixing saidhydrophilic substance and optionally said pH control agent with water,thereby providing a mixture; and mixing said mixture with said emulsion.